Analisis Genomik Karakteristik Resistensi Asam Lambung dan Garam Empedu serta Aktivitas Hemolitik Pada Lactiplantibacillus plantarum subsp. plantarum Mut-7

ADINA AMY R, Dian Anggraini Suroto, S.T.P., M.P., M.Eng., Ph.D.; Dr. Apt. Bondan Ardiningtyas, M.Sc.

2023 | Skripsi | S1 TEKNOLOGI PANGAN DAN HASIL PERTANIAN

Lactiplantibacillus plantarum subsp. plantarum Mut-7 merupakan strain kandidat probiotik indigenus Indonesia yang diisolasi dari Gatot atau makanan fermentasi singkong. Telah diteliti bahwa Lactiplantibacillus plantarum subsp. plantarum Mut-7 memiliki karakteristik probiotik tetapi belum dilakukan konfirmasi sifat resistensi strain Mut-7 terhadap asam lambung dan garam empedu serta ketiadaan aktivitas hemolitik pada Lactiplantibacillus plantarum subsp. plantarum Mut-7 secara genomik. FAO/WHO (2022) dan BPOM (2022) menetapkan bahwa karakterisasi suatu strain probiotik harus mencakup fenotipik dan genomik. Oleh karena itu, penelitian ini dilakukan untuk mengonfirmasi sifat resistensi Lactiplantibacillus plantarum subsp. plantarum Mut-7 terhadap asam lambung dan garam empedu serta aktivitas hemolitik secara genomik. Dilakukan eksplorasi subsistem Lactiplantibacillus plantarum subsp. plantarum Mut-7 untuk mendapatkan sekuens gen protein spesifik dan dilakukan metadata analisis untuk mengetahui keaktifan dari protein spesifik tersebut. Hasil analisis menunjukkan bahwa Lactiplantibacillus plantarum subsp. plantarum Mut-7 memiliki gen protein bile salt hydrolase yang aktif, mengonfirmasi sifat resistensi terhadap garam empedu. Didapatkan pula gen protein dlt operon aktif, berperan dalam D-alanyl-lipoteichoic-acid biosintesis mengonfirmasi sifat resistensi terhadap asam lambung. Lactiplantibacillus plantarum subsp. plantarum Mut-7 memiliki gen protein hemolysin III (hly-III) yang termasuk dalam kelompok protein hemolysin yang diproduksi oleh Bacillus cereus. Berdasarkan hasil eksplorasi gen, diketahui bahwa Lactiplantibacillus plantarum subsp. plantarum Mut-7 tersebut tidak memiliki gen yang berperan dalam memberikan sifat translokasi menyebabkan ketiadaan aktivitas hemolitik.

Lactiplantibacillus plantarum subsp. plantarum Mut-7 is an Indonesian indigenous probiotic strain candicate isolated from Gatot or fermented cassava food. The probiotic properties of Lactiplantibacillus plantarum subsp. plantarum Mut-7 have been researched, although the Mut-7 strain's resilience to gastric acid and bile salts, as well as the lack of hemolytic activity on its genome, have not been proven. The phenotypic and genetic characterization of a probiotic strain is required by FAO/WHO (2022) and BPOM (2022). Therefore, the purpose of this research was to verify Lactiplantibacillus plantarum subsp. plantarum Mut-7's genomic capabilities for resistance to bile salts, stomach acid, and hemolysis. Exploration of the subsystem Lactiplantibacillus plantarum subsp. plantarum Mut-7 was done to obtain specific protein gene sequences and metadata analysis was carried out to determine the activity of these specific proteins. According to the analysis's findings, the bile salt hydrolase protein gene is active in Lactiplantibacillus plantarum subsp. plantarum Mut-7, indicating that it is resistant to bile salts. Additionally, it was discovered that the protein gene in the active operon dlt contributes to the synthesis of D-alanyl-lipoteichoic acid, confirming the organism's resistance to stomach acid. Hemolysin III (hly-III), a protein gene generated by Lactiplantibacillus plantarum subsp. plantarum Mut-7, is a member of the family of hemolysin proteins made by Bacillus cereus. Based on the findings of the gene exploration, it is known that Lactiplantibacillus plantarum subsp. plantarum Mut-7 does not possess a gene involved in supplying translocation capabilities, which leads in the lack of hemolytic activity.

Kata Kunci : Lactiplantibacillus plantarum subsp. plantarum Mut-7, Probiotik, Bile Salt Hydrolase, Resistensi Asam, Hemolitik